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Rabbit Recombinant Monoclonal LCK antibody. Carrier free. Suitable for IP, WB, IHC-P, ICC/IF, Flow Cyt (Intra) and reacts with Human samples.

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Images

Immunoprecipitation - Anti-Lck antibody [Y123] - BSA and Azide free (AB247239), expandable thumbnail
  • Western blot - Anti-Lck antibody [Y123] - BSA and Azide free (AB247239), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-Lck antibody [Y123] - BSA and Azide free (AB247239), expandable thumbnail
  • Western blot - Anti-Lck antibody [Y123] - BSA and Azide free (AB247239), expandable thumbnail
  • Western blot - Anti-Lck antibody [Y123] - BSA and Azide free (AB247239), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBIHC-PICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Tested

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Target data

Function

Non-receptor tyrosine-protein kinase that plays an essential role in the selection and maturation of developing T-cells in the thymus and in the function of mature T-cells. Plays a key role in T-cell antigen receptor (TCR)-linked signal transduction pathways. Constitutively associated with the cytoplasmic portions of the CD4 and CD8 surface receptors. Association of the TCR with a peptide antigen-bound MHC complex facilitates the interaction of CD4 and CD8 with MHC class II and class I molecules, respectively, thereby recruiting the associated LCK protein to the vicinity of the TCR/CD3 complex. LCK then phosphorylates tyrosine residues within the immunoreceptor tyrosine-based activation motifs (ITAM) of the cytoplasmic tails of the TCR-gamma chains and CD3 subunits, initiating the TCR/CD3 signaling pathway. Once stimulated, the TCR recruits the tyrosine kinase ZAP70, that becomes phosphorylated and activated by LCK. Following this, a large number of signaling molecules are recruited, ultimately leading to lymphokine production. LCK also contributes to signaling by other receptor molecules. Associates directly with the cytoplasmic tail of CD2, which leads to hyperphosphorylation and activation of LCK. Also plays a role in the IL2 receptor-linked signaling pathway that controls the T-cell proliferative response. Binding of IL2 to its receptor results in increased activity of LCK. Is expressed at all stages of thymocyte development and is required for the regulation of maturation events that are governed by both pre-TCR and mature alpha beta TCR. Phosphorylates other substrates including RUNX3, PTK2B/PYK2, the microtubule-associated protein MAPT, RHOH or TYROBP. Interacts with FYB2 (PubMed:27335501).

Alternative names

Tyrosine-protein kinase Lck, Leukocyte C-terminal Src kinase, Lymphocyte cell-specific protein-tyrosine kinase, Protein YT16, Proto-oncogene Lck, T cell-specific protein-tyrosine kinase, p56-LCK, LSK, LCK

Recommended products

Rabbit Recombinant Monoclonal LCK antibody. Carrier free. Suitable for IP, WB, IHC-P, ICC/IF, Flow Cyt (Intra) and reacts with Human samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free
Yes
Clone number
Y123
Purification technique
Affinity purification Protein A
Specificity

This antibody is specific for human Lck, it does not cross react with any other SRC family members. WB results were negative for the following Mouse and Rat tissues/cell lines: Ms brain, heart, kidney, spleen, C6, Raw264.7, PC12, NIH3T3, Ms thymus and serum.

Epitope
ab247239 reacts with an epitope located in the region before SH3 domain of Lck.
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Notes

ab247239 is the carrier-free version of Anti-Lck antibody [Y123] ab32149.

Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

Lck also known as lymphocyte-specific protein tyrosine kinase plays an important role in T-cell activation. It serves as an enzyme and is integral in the signal transduction of the immune system. Lck has a molecular weight of approximately 56 kDa. It is expressed predominantly in T lymphocytes and is part of the Src family kinases. This protein is important in phosphorylating substrates leading to downstream signaling events that are necessary for the proper immune function.

Biological function summary

Lck initiates and propagates T-cell receptor (TCR) signaling cascades. It interacts with the CD4 and CD8 co-receptors and triggers the activation of other kinases within the cell. Lck as part of a signaling complex associates with proteins such as ZAP-70 and LAT facilitating the assembly of larger molecular machines necessary for immune responses. Its activity is tightly regulated by phosphorylation with key sites being tyrosine 394 and 505.

Pathways

The Lck protein plays an essential role in TCR signaling and immune responses. It actively participates in the Lck signaling pathway setting off cascades that lead to T-cell activation and differentiation. It interacts closely with other proteins like Fyn and Src making sure that the pathway progresses correctly. In conjunction with the immune synapse Lck ensures the transmission of signals that allow the T-cells to respond to external antigens efficiently.

Associated diseases and disorders

Defective Lck activity can contribute to immunodeficiencies and autoimmune diseases. Proper Lck function is linked to conditions like Severe Combined Immunodeficiency (SCID) and lupus. Dysregulation in Lck activity may also affect proteins such as ZAP-70 worsening disease progression. Understanding Lck's participation in these conditions can lead to better therapeutic interventions and aid in developing targeted treatments for immune-related disorders.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

10 product images

  • Immunoprecipitation - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239), expandable thumbnail

    Immunoprecipitation - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239)

    This data was developed using Anti-Lck antibody [Y123] ab32149, the same antibody clone in a different buffer formulation.Anti-Lck antibody [Y123] ab32149 (purified) at 1:20 dilution (2ug) immunoprecipitating Lck in Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate.
    Lane 1 (input): Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10ug
    Lane 2 (+): Anti-Lck antibody [Y123] ab32149 & Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Lck antibody [Y123] ab32149 in Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

    All lanes: Immunoprecipitation - Anti-Lck antibody [Y123] (Anti-Lck antibody [Y123] ab32149)

    Predicted band size: 58 kDa

  • Western blot - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239), expandable thumbnail

    Western blot - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239)

    This data was developed using Anti-Lck antibody [Y123] ab32149, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-Lck antibody [Y123] (Anti-Lck antibody [Y123] ab32149) at 1/5000 dilution

    Lane 1: Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates at 20 µg

    Lane 2: HuT-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysates at 20 µg

    Lane 3: Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysates at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 58 kDa

  • Flow Cytometry (Intracellular) - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239)

    This data was developed using Anti-Lck antibody [Y123] ab32149, the same antibody clone in a different buffer formulation.

    Intracellular Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling Lck with purified Anti-Lck antibody [Y123] ab32149 at 1/40 dilution (10 ug/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilized with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Western blot - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239), expandable thumbnail

    Western blot - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239)

    This data was developed using Anti-Lck antibody [Y123] ab32149, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-Lck antibody [Y123] (Anti-Lck antibody [Y123] ab32149) at 1/1000 dilution

    All lanes: BxPC-3 (Human pancreas adenocarcinoma epithelial cell) whole cell lysates at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 58 kDa

  • Western blot - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239), expandable thumbnail

    Western blot - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239)

    False colour image of Western blot: Anti-Lck antibody [Y123] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-Lck antibody [Y123] ab32149 was shown to bind specifically to Lck. A band was observed at 60 kDa in wild-type Jurkat cell lysates with no signal observed at this size in Lck knockout cell line Human LCK knockout Jurkat cell line ab273855 (knockout cell lysate Human LCK knockout Jurkat cell lysate ab273809). To generate this image, wild-type and Lck knockout Jurkat cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

    All lanes: Western blot - Anti-Lck antibody [Y123] (Anti-Lck antibody [Y123] ab32149) at 1/1000 dilution

    Lane 1: Wild-type Jurkat cell lysate at 20 µg

    Lane 2: Lck knockout Jurkat cell lysate at 20 µg

    Lane 2: Western blot - Human LCK knockout Jurkat cell line (Human LCK knockout Jurkat cell line ab273855)

    Lane 3: Ramos cell lysate at 20 µg

    Lane 4: A549 cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 58 kDa

    Observed band size: 60 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239)

    This data was developed using Anti-Lck antibody [Y123] ab32149, the same antibody clone in a different buffer formulation.Immunocytochemistry/ Immunofluorescence analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling Lck with purified Anti-Lck antibody [Y123] ab32149 at 1:500 dilution (0.8μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239)

    This data was developed using Anti-Lck antibody [Y123] ab32149, the same antibody clone in a different buffer formulation.Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human Hodgkin lymphoma tissue sections labeling Lck with Purified Anti-Lck antibody [Y123] ab32149 at 1:250 dilution (1.5 μg/ml). Heat mediated antigen retrieval was performed using heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239)

    This data was developed using Anti-Lck antibody [Y123] ab32149, the same antibody clone in a different buffer formulation.Unpurified Anti-Lck antibody [Y123] ab32149 at a 1/250 dilution staining Lck in human lymphoma using Immunohistochemistry, Paraffin Embedded Tissue.

  • Flow Cytometry (Intracellular) - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239)

    This data was developed using Anti-Lck antibody [Y123] ab32149, the same antibody clone in a different buffer formulation.Overlay histogram showing Jurkat cells stained with unpurified Anti-Lck antibody [Y123] ab32149 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (Anti-Lck antibody [Y123] ab32149, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Western blot - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239), expandable thumbnail

    Western blot - Anti-Lck antibody [Y123] - BSA and Azide free (ab247239)

    This data was developed using Anti-Lck antibody [Y123] ab32149, the same antibody clone in a different buffer formulation.

    All lanes: Western blot - Anti-Lck antibody [Y123] (Anti-Lck antibody [Y123] ab32149) at 1/1000 dilution

    All lanes: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate

    Predicted band size: 58 kDa

    Observed band size: 58 kDa

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